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Early Predictive Markers of Pre-Eclampsia

a preeclampsia marker and early prediction technology, applied in the field of preeclampsia preeclampsia detection methods and assays, can solve the problems of not widely accepted or accurate preeclampsia prediction methods, relatively non-specific, and maternal death

Inactive Publication Date: 2015-06-04
UNIV LAVAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent text describes a way to measure markers or ratios to determine if a person is at risk for PE with high accuracy and sensitivity.

Problems solved by technology

PE affects approximately 3-5% of all pregnancies and is a leading cause of maternal death in North America and the UK.
There is no widely accepted or accurate method for the early prediction of PE.
Detection of an abnormality of the blood flow to the uterine artery by Doppler ultrasound in women who later develop PE has been of some predictive use but this abnormality has been found to be relatively non-specific and for this reason has not been adopted in routine clinical practice.
Although some plasma / urine biochemical markers have been shown to be abnormal in the disease process, no single marker has proven to be of adequate sensitivity for use as a predictive indicator.
Although, it is true that PE increases the risk of being affected by cardiovascular diseases later in life, PE is not a cardiovascular disease per se.
Although, there is no widely used treatment for PE (other than premature delivery), Chappell et al.
Despite previous encouraging results of antioxidant vitamin trials, vitamin C and E supplementation did not reduce the rate of PE, but increased the risk of fetal loss or perinatal death and preterm pre-labor rupture of membranes in a large Canadian cohort [2].

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Materials and Methods

[0105]Measurement of F2-isoprostanes by HPLC-MS-MS

Materials

[0106]All F2-isoprostanes and prostaglandin isomers, including 8-iso-15(R)-PGF2α, Ent-8-iso-15(S)-PGF2α, 8-iso-PGF2α, Ent-8-iso-PGF2α, 8-iso-PGF2β, 11β-PGF2α, 15(R)-PGF2α, 5-trans-PGF2α, PGF2α, Ent-PGF2α, PGF2β, iPF2α-IV, (±)5-iPF2α-VI, (±)8,12-iso-iPF2α-VI were purchased from Cayman Chemical (Ann Arbor, Mich., USA) as well as deuterated standards 8-iso-PGF2α-d4, PGF2α-d4, iPF2α-IV-d4, iPF2α-VI-d4, (±) 5-iPF2α-VI-d11, and (±)8,12-iso-iPF2α-VI-d11. Butylated hydroxytoluene (BHT) was bought from Sigma-Aldrich (Oakville, ON, Canada) and sodium chloride (ACS grade) was obtained from Laboratoire Mat (Québec, QC, Canada). All other reagents and solvents were HPLC grade and were purchased from VWR International Inc. (Ville Mont-Royal, QC, Canada).

Preparation of Solutions

[0107]A solution called internal standard containing 50 ng / mL of each deuterated analyte (8-iso-PGF2α-d4, PGF2α-d4, iPF2α-IV-d4, iPF2α-VI-d4, (...

example 2

Results

[0116]The detailed structures of commercially available F2-isoprostanes used to develop the described HPLC-MS-MS method in Example 1 are shown in FIG. 2. Deuterated standards shown in FIG. 3 were used to identify and control for the yield of the isoprostanes extraction or other potential biases throughout the whole experimental procedure. Typical chromatograms for F2-isoprostanes of class III, IV and VI obtained from HPLC-MS-MS analysis are showed respectively in FIGS. 4 to 6. The letters in chromatograms correspond to the structures detailed in FIGS. 2 and 3. For class III F2-isoprostanes, it was not always possible to separate all isomers distinctly. Isomers A and B co-eluted in the same peak on the chromatogram of FIG. 4A. The same phenomenon occurred for isomer C and D and finally I, J and K. The latter indicates that the measurements of commonly studied 8-iso-(15R)-PGF2α and 8-iso-PGF2α are possibly inaccurate because of the co-elution as shown here (FIG. 4A). Moreover, ...

example 3

Improved Separation of Isoprostanes

[0119]We have recently improved the separation of isoprostanes of class VI using a newly developed ion mobility technique. Ion mobility mass spectrometry

[0120]After the chromatographic separation of the isoprostanes as described previously [ref. 8], the samples were introduced in a AB / SCIEX QTRAP 6500 LC / MS / MS System equipped with a SelexION, ion mobility device. The parameters were optimized for each class VI F2-isoprostanes. A high concentration (3.0%) of 2-propanol was used as the differential mobility spectrometer (DMS) chemical modifier. The other operating parameters were set as follows: DMS temperature=300° C. (high), DMS offset=3.0 V, DMS resolution enhancement=low (22 psi) and separation voltage=3750 V. According to those parameters, the optimal compensation voltage was −13.75 V for iPF2α-VI and −10.62 V for both 5-iPF2α-VI and (±)5-8,12-iso-iPF2α-VI (FIG. 7B). This additional step brings a new dimension of separation to the traditional HP...

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Abstract

The present invention relates to a method for measuring levels of class VI isoprostane, an early predictive marker of pre-eclampsia (PE). The present invention also relates to an assay and diagnostic kit for performing the method of predicting PE by measuring the level of this marker and optionally establishing a ratio over other markers such as polyunstaturated fatty acids (PUFA) or beta-carotene. In particular, the method determines the level a class VI isoprostane in a blood sample from a pregnant woman prior to the appearance of symptoms of PE.

Description

CROSS REFERENCES TO RELATED APPLICATIONS[0001]This application is a continuation-in-part of PCT Application No. PCT / CA2013 / 000490, filed May 16, 2013 which claims priority to U.S. Provisional Application Ser. No. 61 / 648,151, filed May 17, 2012, entitled “Early Predictive Markers of Pre-Eclampsia”, the entire contents of which are incorporated by reference herewith.FIELD OF THE INVENTION[0002]The present invention relates to a method and assay for predicting pre-eclampsia (PE). The present invention also relates to a kit for performing the assay of predicting PE.BACKGROUND OF THE INVENTION[0003]PE affects approximately 3-5% of all pregnancies and is a leading cause of maternal death in North America and the UK. This disease, or the threat of onset, is the commonest cause of elective premature delivery, accounting for approximately 15% of all premature births. PE is defined according to the guidelines of the International Society for the Study of Hypertension in Pregnancy and includes...

Claims

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Application Information

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IPC IPC(8): G01N33/88G01N33/92
CPCG01N33/88G01N2800/50G01N2800/368G01N33/92G01N2560/00
Inventor BILODEAU, JEAN-FRANCOISJULIEN, PIERRE
Owner UNIV LAVAL